ATP生物发光法检测细菌药敏技术的建立及应用

目的 采用三磷酸腺苷（ATP）-生物发光法建立一种快速检测细菌抗生素药敏的技术体系.方法 将金黄色葡萄球菌和大肠埃希菌两种菌稀释成不同浓度后,分别加入96孔微孔板中,再加入相应的抗生素,并设立空白对照,混匀后放入细菌培养箱中进行培养,不同时间段取出后采用ATP生物发光仪进行检测,计算其发光率判断细菌是否为耐药与敏感.结果 通过优化条件,确定细菌浓度为5 × 10^5cfu/ml,加入抗生素培养生长4h时为检测最佳时间.与纸片扩散法鉴定的结果进行比较,其匹配率达98.5％,两者差异无统计学意义（P＞0.05）;该方法具有良好的重复性,批内、批间CV均＜10％.结论 ATP生物发光法比现有方法缩短＞1d的时间,且操作简便,快速、准确,成本低,可以广泛用于临床标本检测.

Objective To establish a technical system for rapid detection of antimicrobial susceptibility determination of bacteria by ATP- bioluminescence method. Methods The Staphylococcus aureus and Escherichia coli two were diluted into different concentrations, and incubationed in growth medium with and without antibiotics for different time in the 96 well microplates, then ATP bioluminescence assay was repeated to evaluate the bacteria response to antibiotics.According to the luminescence ratio, the bacteria was determined into resistant or sensitive. Results By optimizing the conditions, when the bacterial concentration was 5 ×10^5cfu/ml and incubationed in growth medium with and without antibiotics for 4H was the best time to detect. Compared with the identification results of the disk diffusion method, the matching rate of the two methods was 98.5% and there was no statistically significant difference between them （ P〉0.05 ） ; the ATP bioluminescence technology had good repeatability, intra, inter CV were less than 10%. Conclusion This method can shorten 1 days or more time than the the established methods, and the operation is simple, rapid, accurate, low cost, can be widely used in clinical specimens.